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. 2015 Jun 1;20(6):10110–10121. doi: 10.3390/molecules200610110

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© 2015 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/4.0/).

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The detection of recombinant in the two transformations by PCR amplification. (a) The detection of recombinant in the first transformation by PCR amplification with primers Cm-F/R. M: Trans DNA Marker II. 1: Negative control, the PCR amplification band of Cm^r gene with the genomic DNA of A. limacinum OUC88 as the template. 2: The PCR amplification band of Cm^r gene with the genomic DNA of A. limacinum OUC_CG as the template. 3: Positive control, the PCR amplification band of Cm^r gene with the plasmid pACYCDuet-1 as the template. B: Blank control; (b) The detection of recombinant in the second transformation by PCR amplification with primers 18S+-F/Ppgk-R. M: Trans15K DNA Marker. 1: The PCR amplification band with genomic DNA of A. limacinum OUC88 as the template; 2: The PCR amplification band (3655 bp) of 18S+-loxP-Cm^r-loxP-Ppgk with genomic DNA of A. limacinum OUC_CG as the template; 3: The PCR amplification band (2185 bp) of 18S+-loxP-loxP-Ppgk with genomic DNA of A. limacinum OUC_EG as the template; B: Blank control.